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Biomedical subjects

J M Anderson

Publications and source records attributed to J M Anderson.

At least 289 records · Page 16Linked to original sources

Effect of albumin coating on the in vitro blood compatibility of Dacron arterial prostheses.

A recirculating in vitro perfusion system was used to assess the effect of albumin precoating on the thrombogenicity of Dacron vascular grafts. A complete analysis of platelet activation was carried out, involving platelet count, release, adhesion and aggregation. Fibrin formation was assessed by measuring fibrinogen levels and fibrinopeptide A production; leucocyte interaction was analysed by measuring total leucocyte count as well as an analysis of cell adhesion to the surface by scanning electron microscopy. The platelet count decreased progressively with perfusion time for Dacron until by 30 min, it had declined to 69% +/- 2% of baseline. The platelet count did not, however, change significantly from baseline when albumin-coated Dacron was tested. Release of platelet factor 4 and beta-thromboglobulin at 180 min for Dacron was 37.8 +/- 29.8 times and 66.9 +/- 18.2 times baseline, respectively, while albumin coating caused significantly less (P less than 0.03) platelet release. Albumin coating diminished coagulation activation and fibrinopeptide A formation. The total leucocyte concentration decreased significantly for Dacron by 180 min, while that for albumin-coated Dacron did not change significantly from baseline levels. Albumin coating produced a film-like covering over the Dacron. For Dacron, there were numerous leucocytes and platelets adherent to the surface, whilst cellular deposition was minimal upon the albumin-coated surface. Thus, albumin coating improved the short-term blood compatibility of Dacron by all of the methods employed in this study.

Albumins↗

Characterization of biomedical polymer-adherent macrophages: interleukin 1 generation and scanning electron microscopy studies.

Macrophage activation following attachment to biomedical polymers was studied using two systems of analysis. Supernatants generated by human peripheral blood monocytes cultured on the surface of several different biomedical polymers were evaluated for the presence of the secreted regulatory protein interleukin 1 (IL1). In addition, each cell-polymer culture surface was subjected to scanning electron microscopy for gross morphological evaluation. Results indicate that, although all materials were efficient in the attachment and activation of cells, the panel of polymers showed a differential capacity in attachment and activation of monocytes. Dacron and polyethylene surfaces had a greater density of cells showing morphology indicative of activation, corresponding to elevated levels of IL1 in these cultures. Biomer and polydimethylsiloxane surfaces showed fewer activated cells and had lower IL1 levels in culture. Expanded polytetrafluorethylene resulted in intermediate levels of IL1 and attached cells showing activated morphology.

Biocompatible Materials↗

ZO-1 mRNA and protein expression during tight junction assembly in Caco-2 cells.

We previously identified and characterized ZO-1 as a peripheral membrane protein specifically associated with the cytoplasmic surface of tight junctions. Here we describe the identification of partial cDNA sequences encoding rat and human ZO-1 and their use to study the assembly of tight junctions in the Caco-2 human intestinal epithelial cell line. A rat cDNA was isolated from a lambda-gtll expression library by screening with mAbs. Polyclonal antibodies were raised to cDNA-encoded fusion protein; several properties of these antibodies support this cDNA as encoding ZO-1. Expression of ZO-1 mRNA occurs in the rat and Caco-2 cells with a major transcript of approximately 7.5 kb. To disrupt tight junctions and study the subsequent process of assembly, Caco-2 cells were grown in suspension for 48 h in Ca++/Mg++-free spinner medium during which time they lose cell-cell contacts, become round, and by immunofluorescence microscopy show diffuse and speckled localization of ZO-1. Within hours of replating at confluent density in Ca++/Mg++-containing media, attached cells show discrete localization of ZO-1 at cell-cell contacts. Within 2 d, fully confluent monolayers form, and ZO-1 localizes in a continuous gasket-like fashion circumscribing all cells. ZO-1 mRNA levels are highest in cells in spinner culture, and upon replating rapidly fall and plateau at approximately 10% of initial levels after 2-3 wk in culture. ZO-1 protein levels are lowest in contact-free cells and rise five- to eightfold over the same period. In contrast, mRNA levels for sucrase-isomaltase, an apical membrane hydrolase, increase only after a confluent monolayer forms. Thus, in this model of contact-dependent assembly of the tight junction, there is both a rapid assembly beginning upon cell-cell contact, as well as a long-term modulation involving changes in expression of ZO-1 mRNA and protein levels.

Animals↗

Characterization of a soybean leaf protein that is related to the seed lectin and is increased with pod removal.

Levels of several polypeptides in addition to the vegetative storage protein (VSP) increase in soybean leaves following depodding. Two of these polypeptides interact specifically with antibodies raised against the seed lectins of Phaseolus vulgaris and soybean. The two polypeptides, which had apparent molecular masses of 29,000 daltons and 33,000 daltons, were present in the sink-deprived plants but not in control podded plants and were the subunit polypeptides of a glycoprotein designated lectin-related protein (LRP). Soybean LRP was purified to near homogeneity by a combination of ammonium sulfate precipitation and gel filtration. Dialysis of the resuspended ammonium sulfate precipitate caused LRP to reprecipitate, and LRP was soluble only in the presence of molar NaCl. The native relative molecular mass of LRP was 119,000 daltons, a size consistent with a tetrameric organization of the two polypeptides. LRP precipitated during dialysis in association with a 28,000 dalton polypeptide. The protein coprecipitating with LRP was identified as the dimer of the 28,000 dalton subunit of VSP, one of three native isomeric forms of VSP occurring in leaves of depodded plants. Although the specific association between LRP and VSP was intriguing, an in vivo interaction between LRP and VSP was doubtful. LRP was shown to be immunologically similar to soybean agglutinin but did not have detectable hemagglutinating activity. LRP also was shown to be made up of polypeptides distinct from soybean agglutinin.

Journal Article↗

Managing chronic illness in the family: women as caretakers.

This paper is based on an ethnographic study examining how families caring for a chronically ill child in the home construct their experiences of illness. The role of women caretakers is examined, and it is argued that the term family glosses over the work that women do in caretaking. The 'ideology of competence' which determines women's subjective experiences is discussed, and the notion is forwarded that health professionals rely upon this ideology to get the job of caretaking done in the home. Some implications of this discussion for health care delivery are presented.

Canada↗

Implementation of a self-health project by baccalaureate students.

With increasing evidence that life-style is an important influence on health, three nursing faculty members at the University of Minnesota implemented a learning project to enable students to assess, plan, and evaluate their own life-styles. The goal was to have the students attempt to make positive changes. As part of a "health concepts" nursing course, students became much more aware of social, economic, environmental, and cultural factors that either enhanced or detracted from their ability to achieve their ideal life-styles. The students responded favorably to this assignment because of the potential benefits of investing in themselves while pursuing the rigorous program leading to a nursing degree.

Education, Nursing, Baccalaureate↗

Chlamydia trachomatis infection in infants: a prospective study.

In a prospective study of Chlamydia trachomatis infection in pregnancy, 198 mothers positive for chlamydial antigen were identified; the infants of 174 were followed for up to six months and C trachomatis was recovered in cell culture from 43 infants (25%). Conjunctivitis occurred in significantly more infants who were positive for C trachomatis (20 of 43, 47%) than in those who were negative (18 of 131, 14%). There were also significantly more lower respiratory tract infections among infants with positive cultures (six of 43, 14%, compared with three of 131, 2%). The chlamydial antigen enzyme linked immunosorbent assay (ELISA) was positive in 61 of 131 infants from whom C trachomatis was not recovered in cell culture. False positive results were usually associated with the isolation of Staphylococcus aureus from samples of pharyngeal aspirate. Our results confirm that C trachomatis infection is a common cause of neonatal conjunctivitis, and respiratory infection in the first few months of life, with an incidence of 8.2/1000 live births. Because the infection is easily treated by oral erythromycin, however, screening during pregnancy is not warranted.

Chlamydia Infections↗

Observer variability in histopathological reporting of cervical biopsy specimens.

Sections from 100 cervical biopsy specimens were studied by 12 consultant histopathologists to determine the robustness of the existing pathology terminology and classification. Analysis by kappa statistics showed good agreement in the diagnosis of CIN 3 and squamous carcinoma but an inability to distinguish accurately between the lesser grades of CIN. It is recommended that the classification be changed to low grade (present CIN 1 and 2) and high grade (present CIN 3) categories alone. There was very poor agreement in the identification of cellular changes associated with human papilloma virus (HPV) infection.

Biopsy↗

ZO-1 and cingulin: tight junction proteins with distinct identities and localizations.

The relative localization of ZO-1 and cingulin, the only two known components of the tight junction, was compared in Madin-Darby canine kidney (MDCK) cells, chicken small intestine, rat kidney distal convoluted tubule, and a hepatoma cell line. Immunoblot analysis demonstrated that cingulin and ZO-1 are immunologically unrelated and that, in the colon, cingulin is a single polypeptide with a molecular mass of 140 kDa. Immunofluorescent localization of cingulin and ZO-1 in confluent monolayers of MDCK cells showed identical staining patterns. However, subconfluent MDCK cells showed distinct localizations of the two proteins. Both cingulin and ZO-1 were found at the plasma membrane only at areas of cell-cell contact, but cingulin was diffusely distributed within the cytoplasm, whereas ZO-1 showed a more clustered internal arrangement. Cingulin and ZO-1 were identically localized at the plasma membrane of hepatoma tissue culture (HTC) cells at sites of cell-cell contact. In chicken intestine examined at the ultrastructural level, immunogold particles associated with cingulin were found approximately three times farther from the junctional membrane than those affiliated with ZO-1.

Animals↗

Hepatic immunohistochemical localization of the tight junction protein ZO-1 in rat models of cholestasis.

Structural alterations in hepatocyte tight junctions accompanying cholestasis were investigated using immunolocalization of ZO-1, the first known protein component of the tight junction. Disruption in the paracellular barrier function of the tight junction has been proposed to allow reflux of bile into the blood. Cholestasis was induced in 210 to 235 g male Sprague-Dawley rats either by five consecutive daily subcutaneous injections of 17-alpha-ethinyl estradiol (0.5 mg/kg/d in propylene glycol) or ligation of the common bile duct for 72 hours. The structural organization of the tight junction was assessed in each model by indirect immunofluorescent and immunoperoxidase staining for ZO-1 on frozen sections of liver and compared with controls. In control, sham-operated, and estradiol-injected animals, ZO-1 localizes in a uniform continuous manner along the margins of the canaliculi. In contrast, bile duct ligation results in the appearance of numerous discontinuities in ZO-1 staining accompanied by dilation or collapse of the lumenal space. Tissue content of the ZO-1 protein, as determined by quantitative immunoblotting, was unaffected in either cholestatic model compared with controls. These findings indicate that the molecular organization of the tight junction can be assessed from immunostaining patterns of ZO-1 in frozen sections of cholestatic livers. Under these experimental conditions, the organization of the tight junction at the level of the ZO-1 protein is altered by bile duct obstruction but not by ethinyl estradiol.

Animals↗

Human monocyte/macrophage activation and interleukin 1 generation by biomedical polymers.

In vitro cell culture techniques were used to evaluate the effect of several clinically significant biomedical polymers on monocyte activation and Interleukin 1 (IL1) production. Isolated human peripheral blood monocytes were cultured in the presence of a panel of five biomedical polymers routinely used in a variety of clinical applications: Polyethylene (PE), silica-free poly-dimethylsiloxane (PDMS), woven Dacron fabric, expanded polytetrafluoroethylene (ePTFE) and the segmented polyurethane, Biomer. Monocytes generated IL1 in the presence of all five materials. Maximal levels of IL1 were generated at 24 h in monocyte-polymer cultures supplemented with serum and additionally stimulated with lipopolysaccharide (LPS). No difference was observed due to serum source. Results from cultures supplemented with fetal bovine serum were not significantly different from those obtained with human serum supplemented cultures. The thymocyte proliferative activity generated by monocytes in the presence of these biomedical polymers was neutralized by a specific polyclonal anti-IL1 antiserum. Statistically significant differences in IL1 production were observed between polymers, allowing their classification according to reactivity into high (Dacron, PE), intermediate (ePTFE) and low (Biomer, PDMS) reactive groups.

Biocompatible Materials↗

The epithelial tight junction: structure, function and preliminary biochemical characterization.

The tight junction, or zonula occludens (ZO), forms a semi-permeable barrier in the paracellular pathway in most vertebrate epithelia. The ZO is the apical-most member of a series of intercellular junctions, collectively known as the junctional complex, found at the interface of the apical and lateral cell surface. This structure not only restricts movement of substances around the cells, but may also serve as a 'fence' acting to maintain the cell surface compositional polarity characteristic of epithelial cells. The morphology and physiology of the ZO have been well documented and are briefly reviewed here. The biochemistry of this important intercellular junction remains largely unknown, although a tight junction-specific polypeptide called 'ZO-1' has recently been identified. Preliminary observations regarding the role of this peripheral phosphoprotein in the biology of the ZO are presented.

Animals↗

An assessment of colostomy irrigation.

One hundred patients with permanent sigmoid colostomies were surveyed to determine their satisfaction and success with the "irrigation" technique of colostomy management. Most patients who irrigate their colostomies achieve continence. Odors and skin irritation are minimized. The irrigation method is economical, time efficient, and allows a reasonably liberal diet. It avoids bulky appliances and is safe. In appropriately selected patients, the irrigation technique is the method of choice for management of an end-sigmoid colostomy.

Adult↗

In vitro and in vivo interactions of cells with biomaterials.

The biocompatibility of materials at an implant site involves a complex interaction of cells and tissues with the biomaterial. This cell-cell and cell-polymer interaction evokes the release of mediators such as chemotactic and growth factors that elicit and sustain inflammatory responses at the implant site. In this review, we summarize the interaction of cells with biomaterials in vitro and in vivo.

Animals↗

In vivo biocompatibility of catheter materials.

The inflammatory response to four different catheter materials was quantitatively characterized using an in vivo cage implant system over a 21 d implantation period. The greatest differences between materials were observed at day 4, where the total leucocyte concentration in the exudate decreased in the following order: AE-PVC greater than A-PU greater than D-PU1 greater than D-PU2. Similar trends were observed for the polymorphonuclear leucocyte (PMN) and macrophage concentrations at day 4. At day 7, this trend continued, but by day 21 the total leucocyte concentration, PMN concentration and macrophage concentration had decreased to comparable values for all materials. The extracellular alkaline phosphatase activity at day 4 was lowest for the AE-PVC exudate samples. Similar results were observed for the extracellular acid phosphatase activity but after that time point no significant differences were observed. From these results, it appears that AE-PVC is least biocompatible compared with the other three catheter materials, which were polyurethanes.

Acid Phosphatase↗

Human platelet interactions with surfaces of type I collagen, chondroitin-4-sulphate, and chondroitin-6-sulphate in vitro.

In vitro human platelet interactions with surfaces of type I collagen, chondroitin-6-sulphate (CH-6-S), chondroitin-4-sulphate (CH-4-S), a CH-6-S/collagen layer, and a collagen-CH-6-S complex were investigated. Polystyrene and silanized glass served as controls. Platelet counts, platelet factor 4 released, and platelet aggregating ability for the different surfaces were compared with controls. Platelet count and platelet factor 4 release data showed that there were no differences between surfaces of CH-6-S, CH-4-S and the controls. However, significant differences in platelet counts and platelet factor 4 released were found when collagen, the CH-6-S/collagen layer, and the collagen-CH-6-S complex were compared with controls. The pure type I collagen surface had the greatest influence on platelet activation. The collagen-CH-6-S complex had a greater effect that the CH-6-S/collagen layer on platelet activation. It appears that chondroitin-6-sulphate can modify the platelet activity of type I collagen.

Blood Platelets↗

Characterization of ZO-1, a protein component of the tight junction from mouse liver and Madin-Darby canine kidney cells.

ZO-1, originally identified by mAb techniques, is the first protein shown to be specifically associated with the tight junction. Here we describe and compare the physical characteristics of ZO-1 from mouse liver and the Madin-Darby canine kidney (MDCK) epithelial cell line. The ZO-1 polypeptide has an apparent size of 225 kD in mouse tissues and 210 kD in canine-derived MDCK cells as determined by SDS-PAGE/immunoblot analysis. ZO-1 from both sources is optimally solubilized from isolated plasma membranes by either 6 M urea or high pH conditions; partial solubilization occurs with 0.3 M KCl. The nonionic detergents, Triton X-100 and octyl-beta-D-glucopyranoside, do not solubilize ZO-1. These solubility properties indicate that ZO-1 is a peripherally associated membrane protein. ZO-1 was purified to electrophoretic homogeneity from [35S]methionine metabolically labeled MDCK cells by a combination of gel filtration and immunoaffinity chromatography. Purified ZO-1 has an s20,w of 5.3 and Stokes radius of 8.6 nm. These values suggest that purified ZO-1 is an asymmetric monomeric molecule. Corresponding values for mouse liver ZO-1, characterized in impure protein extracts, were 6 s20,w and 9 nm. ZO-1 was shown to be a phosphoprotein in MDCK cells metabolically labeled with [32P]orthophosphate; analysis of phosphoamino acids from purified ZO-1 revealed only phosphoserine. ZO-1 epitope number was determined by Scatchard analysis of competitive and saturable binding of two different 125I-mAbs to SDS-solubilized proteins from liver and MDCK cells immobilized on nitrocellulose. Saturation binding occurs at 26 ng mAb/mg liver and 63 ng/mg of MDCK cell protein. This is equivalent to 30,000 ZO-1 molecules per MDCK cell assuming a single epitope/ZO-1 molecule.

Animals↗